Quinoline- and naphthalenecarboxamides, pharmaceutical compositions and methods of inhibiting calpain

ABSTRACT

Pharmaceutical compositions and methods of inhibiting calpain using novel quinoline- or naphthatenecarboxamides are disclosed.

This application is a 371 of PCT/US98/04874 filed Mar. 13, 1998 whichclaims the benefit of priority to this provisional application,60/040,583, filed Mar. 14, 1997.

SUMMARY OF THE INVENTION

This invention relates to novel chemical compounds which are quinoline-or aphthalenecarboxamides. The claimed pharmaceutical compositions andmethods use those compounds as active ingredients to inhibit calpain andthus are useful in the treatment of, for example, neurodegenerativedisorders, strokes and traumatic brain injury.

BACKGROUND OF THE INVENTION

Calpains are calcium--dependent cysteine proteases present in a varietyof tissues and cells. Excessive activation of calpain provides amolecular link between ischaemia or injury induced by increases inintraneuronal calcium and pathological neuronal degeneration. If theelevated calcium levels are left uncontrolled, serious structural damageto neurons may result. Recent research has suggested that calpainactivation may represent a final common pathway in many types of braindamage. Selective inhibition of calpain would, therefore, be anattractive therapeutic approach in the treatment of neurodegenerativediseases. Exemplary of these diseases would be myocardial ischaemia,cerebral ischaemia, muscular dystrophy, stroke, Alzheimer's disease, ortraumatic brain injury. The compounds of this invention may also beuseful in the treatment of cataracts and platelet aggregation.

DETAILED DESCRIPTION OF THE INVENTION

The compounds which are the active ingredients of the pharmaceuticalcompositions and methods of this invention are represented by thefollowing formula: ##STR1## in which: X is CH or N;

R is CH₂ Ph, --(CH₂)₃ CH₂ NR₁ R₃, CH₂ CH(CH₃)₂ or CH₂ PhOR₂ ;

R₁ is COOCH₂ Ph, SO₂ CH₃, SO₂ aryl, COOCH_(2pyridyl) (or substitutedpyridyl);

R₂ is H, CH₃, CH₂ Ph or CH_(2pyridyl) ;

R₃ is H, CH₃ or lower alkyl;

Z is CHO, COCH₂ F, COCOOH, COCOOalkyl, COCONHalkyl, COCO(CH₂)_(n) aryl,COCONHCH(R)COOH or COCH₂ O--(3-phenylisoxazol-5-yl);

n is 1 to 6; ##STR2## Y=absent, phenyl, substituted phenyl, pyridyl orsubstituted pyridyl, or a pharmaceutically acceptable salt thereof.

Preferred compounds are those where the stereochemistry at the R groupcorresponds to that of the naturally occurring amino acids. Alsopreferred are those compounds where X is N, Z is CHO and R is CH₂ Ph or--(CH₂)₃ CH₂ NR₁ R₃.

The following preferred compounds are representative of the compounds ofthe invention:

(S)-N-(1-formyl-2-phenylethyl)-2-phenyl-4-quinolinecarboxamide

(S)-2-(4-chlorophenyl)-N-(1-formyl-2-phenylethyl)-4-quinolinecarboxarmide

(S)-2-[1,1'-biphenyl]-4-yl-N-(1-formyl-2-phenylethyl)-4-quinolinecarboxamide

(S)-2-(1-adamantyl)-N-(1-formyl-2-phenylethyl)-4-quinolinecarboxamide

(S)-N-(1-formyl-2-phenylethyl)-2-(4-phenoxyphenyl)4-quinolinecarboxamide

(S)-2-[1,1'-biphenyl]-2-yl-N-(1-formyl-2-phenylethyl)-4-quinolinecarboxamide

(S)-N-(1-formyl-2-phenylethyl)-2-(2-pyridinylethynyl)-4-quinolinecarboxamide

(S)-N-(1-formyl-2-phenylethyl)-2-(4-phenyl-1-piperazinyl)-4-quinolinecarboxamide

(S)-N-[1-formyl-2-phenylethyl]-2-[(3-(pyridinyl)-4-phenyl]-4-quinolinecarboxamide

(S)-N-[1-formyl-5-[(phenylsulfonyl)amino]pentyl]-2-phenyl-4-quinolinecarboxamide

(S)-N-[1-formyl-5-[N'-(carbo-4-pyridinemethyloxy)pentyl]-2-[phenyl-]4-quinoline-carboxamide

(S)-N-(1-formyl-2-phenylethyl)-2-(phenylethynyl)-4-quinolinecarboxamide

N-[3-(n-butylamino)-2,3-dioxo-1-(phenylmethyl)]-2-(phenylethynyl)-4-quinoline-carboxamide.

Compounds of Formula I where X is N are prepared by the methodsdescribed in Schemes 1-4. ##STR3##

The 2-substituted quinolines 1 where Q-Y is phenyl or para-biphenyl areavailable from Aldrich Chemical Company. 1 is converted to the amidealcohol 2 by standard coupling conditions[(S)-(-)-2-amino-3-phenyl-1-propanol,benzotriazol-1-yloxytris(dimethylamino)-phosphoniumhexafluorophosphate(BOP), triethylamine, methylene chloride)]. The amide alcohol may bepurified by silica chromatography. Oxidation of 2 (the Dess-Martinreagent in methylene chloride is prefered, but not limiting) affords thealdehyde 3. This procedure can be repeated with a wide variety of2-substituted quinoline-4carboxylates and with a wide variety of aminoalcohol derivatives. Those derived from the naturally occurring aminoacids are preferred.

Compounds of Formula I wherein the quinoline containing the desiredsubstituent at C-2 is not commercially available are prepared by themethods described in Schemes 2-4. ##STR4##

2-Chloroquinoline-4-carboxylic acid, 4, is available from ICN ChemicalCompany. 4 is converted to the amide alcohol 5 by standard couplingconditions as in Scheme 1 [(S)-(-)-2-amino-3-phenyl-1-propanol,benzotriazol-1-yloxytris(dimethylamino)-phosphoniumhexafluorophosphate(BOP), triethylarnine, methylene chloride)]. The chloro substituent of 5is then replaced with the desired Q-Y group by palladium catalyzedcoupling chemistry (for acetylene coupling see Sakamoto et al., Chem.Pharm. Bull., 1984, 32, 4666-4669; for boronic acid coupling see Finchet al., J. Chem. Soc. Perkin I, 1994, 9, 1193-1203). In this way, theuse of substituted acetylenes (2-pyridylacetylene) and boronic acidderivatives (2-phenylphenylboronic acid) are added to the C-2 positionof the quinoline ring. The amide alcohol 2 may be purified by silicachromatography. Oxidation of 2 (the Dess-Martin reagent in methylenechloride is prefered, but not limiting) affords the aldehyde 3. Thisprocedure can be repeated with a wide variety of amino alcoholderivatives. Those derived from the naturally occurring amino acids arepreferred.

Compounds of Formula I wherein variations at the C-2 quinolinesubstituent, in addition to the method described in Scheme 2, aredescribed in Scheme 3. ##STR5##

Compound 6 is prepared according to the boronic acid coupling methoddescribed in Scheme 2. Tin mediated coupling of 6 provides the amidealcohol 7 which on oxidation (preferably with Dess-Martin reagent)provides the aldehyde 8.

Additional variation at C-2 of the quinoline ring is accomplished asdemonstrated in Scheme 4. ##STR6##

5 is obtained as described in Scheme 2. Treatment with a nucleophiliccompound such as N-phenylpiperazine provides the amide alcohol 9 whichis conerted into the aldehyde 10 on oxidation (preferably with theDess-Martin reagent). This method is versatile in that a wide variety ofamines and other nucleophilic species can be used to displace thequinoline C-2 chloro group.

Variation of the side chain is accomplished using the functionality ofamino acid side chains as demonstrated in Scheme 5. ##STR7##

Compound 12 is obtained by BOP coupling as previously described (e.g.,Scheme 1). Removal of the BOC protecting group using acidic conditionsaffords 13. The amino group can be reacted in different ways to afford adiversity of products. For example, reacting with a sulfonyl chloridesuch as phenylsulfonyl chloride affords 14, which upon reduction to 15(R' is C₆ H₅ SO₂ NH) followed by oxidation provides the desired product16 (R' is C₆ H₅ SO₂ NH). Alternatively, the amino group of 13 can beconverted into an intermediate isocyanate group (phosgene, pyridine, CH₂Cl₂) such as 14 (R' is --N═C═O) which upon reaction with an appropriatenucleophilic substrate such as an alcohol (HOR), an amine or amercaptan, affords the corresponding product 14 (R'=--NHCOOR). In such areaction, treatment of 14 (R' is --N═C═O) with 4-pyridinecarbinolproduces 14 (R' is --NHCOOCH₂ -4-pyridine). Reduction of the esteraffords 15 (R' is --NHCOOCH₂ -4-pyridine) and oxidation provides thedesired product 16 (R' is --NHCOOCH₂ -4-pyridine).

Although these methods illustrate the preparation of compounds for whichZ=CHO, alternative "enzyme reactive groups" can be substituted as hasbeen extensively described in the literature (J. Med. Chem., 1994, 37,2918-2929, J. Med. Chem., 1993, 36, 3472-3480, J. Med. Chem. 1990, 33,11-13, Biochem. J., 1986, 239, 633-640, J. Med. Chem., 1992, 35,216-220). In addition, these methods are not intended to limit the scopeof the possible R groups which can be readily derived from any aminoalcohol or amino acid by methods well known in the art.

Also included in the scope of the present invention are pharmaceuticallyacceptable salts of the compounds of Formula I. Preferred salts include,but are not limited to, hydrochloride, hydrobromide, citrate, tartrate,malate, maleate, lactate, gluctose, 1,6-diphosphate, phosphate,succinate, sulfate, aspartate, adipate, methanesulfonate, laurylsulfate, diguaiacyl phosphate, diacetyl sulfate, glutamate, edetate,ethylene diamine, sodium, potassium, calcium and ethanolamine salts.Such salts are prepared according to standard procedures well known inthe art.

The pharmaceutical activity of the compounds of this invention isdemonstrated by inhibition of calpain in vitro by the assay proceduredescribed by Sasaki et al., J. Biol. Chem. 1984, 259, 12489-12494. Theassays were performed using synthetic fluorogenic substrates. Inhibitionof enzyme activity was calculated on the percent decease in the rate ofsubstrate hydrolysis in the presence of inhibitor relative to the ratein its absence. IC₅₀ s(nM) were calculated. Table 1 demonstrates theresults of testing representative compounds of Formula I.

                                      TABLE 1                                     __________________________________________________________________________     ##STR8##                                                                     Z      R              Q-Y            IC.sub.50 (mM)                           __________________________________________________________________________    CHO    CH.sub.2 Ph    Ph             65                                       CHO    CH.sub.2 Ph    Ph-4-Cl        66                                       CHO    CH.sub.2 Ph    Ph-4-Ph        20                                       CHO    CH.sub.2 Ph    1-adamantyl    123                                      CHO    CH.sub.2 Ph    Ph-4-OPh       32                                       CHO    CH.sub.2 Ph    Ph-2-Ph        19                                       CHO    CH.sub.2 Ph                                                                                   ##STR9##      16                                       CHO    CH.sub.2 Ph                                                                                   ##STR10##     38                                       CHO    CH.sub.2 Ph                                                                                   ##STR11##     46                                       CHO    (CH.sub.2).sub.3 CH.sub.2 NHSO.sub.2 Ph                                                      Ph             42                                       CHO    (CH.sub.2).sub.3 CH.sub.2 NHCO.sub.2 CH.sub.2 -4-pyridyl                                     Ph             160                                      CHO    CH.sub.2 Ph                                                                                   ##STR12##     12                                       COCONHn-Bu                                                                           CH.sub.2 Ph                                                                                   ##STR13##     77                                       __________________________________________________________________________

The above results clearly indicate that all compounds tested exhibitedsignificant inhibition of calpain.

The pharmaceutical compositions of this invention employed to inhibitcalpain comprise a pharmaceutical carrier and as the active ingredient acompound of Formula I. The active ingredient will be present in thecompositions of this invention in an effective amount to inhibitcalpain. Preferably, the compositions contain the active ingredient ofFormula I in an amount of from about 0.1 mg to about 250 mg,advantageously from about 25 mg to about 150 mg per dosage unit.

The pharmaceutical carrier may be, for example, a solid or liquid.Exemplary of solid carriers are lactose, magnesium stearate, sucrose,talc, stearic acid, gelatin, agar or acacia. Exemplary of liquidcarriers are syrups, peanut oil, olive oil, propylene glycol,polyethylene glycol and water.

A wide variety of pharmaceutical forms may be employed. Thus, if a solidcarrier is used, the preparation can be tabletted or placed in a hardgelatin capsule. If a liquid carrier is used, the preparation may be inthe form of a soft gelatin capsule, placed in an ampule, a liquidsuspension, syrup or suspension.

Preferably, parenteral solutions or suspensions are employed. Theycomprise the active compound in a sterile aqueous or oil carrier suchas, for example, peanut oil, polyethylene glycol or polyvinylpyrolidone. Preferably, such solutions contain the active compound inthe range of 0.1 to 140 mg/kg of body weight of the patient to whom itwill be administered. The sterile parenteral solutions may also containadditives such as, for example, preservatives such as benzyl alcohol andbuffering agents to bring the injectable preparation to a satisfactorypH. Stabilizing agents such as ascorbic acid or sodium bisulfate mayalso be employed. DMSO or alcoholic solvents may be used to aid in thesolubility and penetration of the calpain inhibitor.

The sterile aqueous solutions can also be lyophilized and reconstitutedprior to administration.

The parenteral solution may be administered subcutaneously,intravenously, intramuscularly, interperitoneally, intrastemally or byintrathecal injection directly into the central nervous system.

The pharmaceutical compositions are prepared by conventional techniquesinvolving procedures such as mixing, granulating and compressing todissolve the ingredients as appropriate to the desired preparation.

The method of inhibiting calpain according to this invention comprisesadministering to an animal or human in an amount sufficient to inhibitcalpain a compound of Formula I.

Preferably the compounds of Formula I are administered in conventionaldosage unit forms prepared by combining an appropriate dose of thecompound with standard pharmaceutical carriers.

Most preferably, the active ingredients of Formula I will beadministered in a daily dosage regimen of from about 2.0 mg to about 1.0g, most preferably from about 50 mg to about 400 mg. Advantageously,equal doses will be administered two to four times a day. When theadministration is carried out as described above, inhibition of calpainis produced.

The route of administration of the pharmaceutical compositions of thisinvention and in accordance with the methods of this invention isinternal, more specifically either oral or preferably parenteral, in anamount sufficient to produce the desired biological activity.

The following examples are not limiting but are illustrative of thecompounds and compositions of this invention and the process for theirpreparation.

EXAMPLE 1 Preparation of(S)-N-(1-formyl-2-phenylethyl)-2-phenyl-4-quinolinecarboxamide

(a)(S)-N-(1-Hydroxymethyl-2-phenylethyl)-2-phenyl-4-quinoline-carboxamide

To a solution of 2-phenyl-4-quinoline-carboxylic acid (0.33 g, 1.3 mmol,Aldrich Chemical Company) in methylene chloride (5 mL) was addedbenzotriazol-1-yloxytris-(dimethyamino)phosphoniumhexafluorophosphate(BOP) (0.63 g, 1.43 mmol). The resulting mixture was shaken at roomtemperature for 5 min. (S)-(-)-2-arnino-3-phenyl-1-propanol (0.2 g, 1.3mmol) was added along with triethylamine (0.2 mL, 1.43 mmol). Theresulting mixture was shaken at room temperature for 24 h. Methylenechloride (10 mL) was added and the organic layer was washed with NaHCO₃,H₂ O, brine, dried (MgSO₄), filtered, and concentrated in vacuo to givean oil. The oil was purified by flash chromatography (silica gel, 30-80%EtOAc/hexane) to yield the title compound as a white solid (0.3 g, 60%).MS (ES+) m/e 383.5 [M+H]⁺, 405 [M+Na]⁺, 787 [2M+Na]⁺.

(b) (S)-N-(1-Formyl-2-phenylethyl)-2-phenyl-4-quinolinecarboxamide

To a solution of the compound of Example 1(a) (0.1 g, 0.26 mmol)dissolved in methylene chloride (5 mL) was added1,1,1-triacetoxy-1,1-dihydro-1,2-benziodoxol-3(1H)-one (Dess-Martinperiodinane) (0.12 g, 0.3 mmol). The resulting mixture was shaken atroom temperature for 1 hr. Sodium thiosulfate solution (10%) (2 mL) wasadded and the mixture was shaken for 10 min. The organic layer waswashed with NaHCO₃, brine, dried (MgSO₄), filtered, and concentrated invacuo to give a tan solid. The solid was recrystallized from diethylether to yield the title compound as a pale white solid (0.053 g, 54%).¹ H NMR (400 MHz, CDCl₃) δ 9.83 (s, 1H), 8.09 (m, 3H), 7.78 (m, 2H),7.57 (m, 4H), 7.31 (m, 6H), 6.6 (d, 1H), 5.15 (t, 1H), 3.5 (d, 2H). MS(ES+) m/e 381.4 [M+H]⁺, 413.4 [M+H+CH₃ OH]⁺.

EXAMPLE 2 Preparation of(S)-2-(4-Chlorophenyl)-N-(1-formyl-2-phenylethyl)-4-quinolinecarboxamide

Following the procedures of Examples 1(a), and 1(b) except substituting2-phenyl-4-quinoline-carboxylic acid with2-(4-chlorophenyl)-4-quinoline-carboxylic acid, the title compound wasprepared as a cream solid (0.06 g, 50% for two steps). MS (ES+) m/e 415[M+H]⁺.

EXAMPLE 3 Preparation of(S)-2-[1.1'-Biphenyl]-4-yl-N-(1-formyl-2-phenylethyl)-4-quinolinecarboxamide

Following the procedures of Examples 1(a), and 1(b) except substituting2-phenyl-4-quinoline-carboxylic acid with2-[1,1'-biphenyl)-4-quinoline-carboxylic acid, the title compound wasprepared as a yellow solid (0.07 g, 55% for two steps). MS (ES+) m/e 457[M+H]⁺.

EXAMPLE 4 Preparation of(S)-2-(1-Adamantyl)-N-(1-formyl-2-phenylethyl)-4-quinolinecarboxamide

Following the procedures of Examples 1(a), and 1(b) except substituting2-phenyl-4-quinoline-carboxylic acid with2-(1-adamantyl)-4-quinoline-carboxylic acid, the title compound wasprepared as a white solid (0.06 g, 50% for two steps). MS (ES+) m/e 439[M+H]⁺, 471 [M+H+CH₃ OH]⁺.

EXAMPLE 5 Preparation of(S)-N-(1-Formyl-2-phenylethyl)-2-(4-phenoxyphenyl)-4-quinolinecarboxamide

(a)(S)-N-([1-Hydroxymethyl-2-phenylethyl)-2-chloro-4-quinolinecarboxamide

Following the procedure of Example 1(a) except substituting2-phenyl-4-quinolinecarboxylic acid with 2-chloro-4quinoline-carboxylicacid, the title compound was prepared as a white solid (0.06 g, 70%). MS(ES+) m/e 341 [M+H]⁺.

(b)(S)-N-(1-Hydroxymethyl-2-phenylethyl)-2-(4-phenoxyphenyl)-4-quinoline-carboxamide

To a solution of the compound of Example 5(a) (0.15 g, 0.44 mmol) in drytoluene (4 mL) under argon atmosphere was addedtetrakis(triphenylphosphine)-palladium(0) (25 mg, 0.022 mmol) followedby 4-phenoxyphenylboronic acid (0.18 g, 0.88 mmol; Sigma ChemicalCompany), sodium carbonate (2M solution in H₂ O, 0.6 mL) and ethanol (2mL). The resulting mixture was refluxed for 4 h. Methylene chloride (50mL) was added and the organic layer was washed with H₂ O, brine, dried(MgSO₄), filtered, and concentrated in vacuo to give a solid. Theresulting crude yellow solid was triturated with methylene chloride (40mL, and filtered to yield the title compound as an off-white solid (0.15g, 75%). MS (ES+) m/e 475 [M+H]⁺.

(c)(S)-N-(1-Formyl-2-phenylethyl)-2-(4-phenoxyphenyl)4-quinolinecarboxamide

Following the procedure of Example 1(b) except substituting the compoundof Example 1(a) with the compound of Example 5(b), the title compoundwas prepared as a light yellow solid (0.1 g, 73%). MS (ES+) m/e 473[M+H]⁺.

EXAMPLE 6 Preparation of(S)-2-[1.1'-Biphenyl]-2-yl-N-(1-formyl-2-phenylethyl)-4-quinolinecarboxamide

(a)(S)-2-[1,1'-Biphenyl]-2-yl-N-(1-hydroxymethyl-2-phenylethyl)-4-quinoline-carboxamide

To a solution of the compound of Example 5(a) (0.15 g, 0.44 mmol) in drytoluene (4 mL) under argon atmosphere was addedtetrakis(triphenylphosphine)-palladium(0) (25 mg, 0.022 mmol) followedby (2-phenyl)phenylboronic acid (0.18 g, 0.88 mmol; synthesizedaccording to the procedure of Kelly et al., J. Amer. Chem. Soc., 1990,112, 8024-8034), sodium carbonate (2M solution in H₂ O, 0.6 mL) andethanol (2 mL). The resulting miture was refluxed for 4 h. Methylenechloride (50 mL) was added and the organic layer was washed with H₂ O,brine, dried (MgSO₄), filtered, and concentrated in vacuo to give afoam. The foam was purified by flash chromatography (silica gel, 25-50%EtOAc/hexane) to yield the title compound as a glassy foam (0.11 g,59%). MS (ES+) m/e 459.3 [M+H]⁺.

(b)(S)-2-[1,1'-Biphenyl]-2-yl-N-(1-formyl-2-phenylethyl)-4-quinolinecarboxamide

Following the procedure of Example 1(b) except substituting the compoundof Example 1(a) with the compound of Example 6(a), the title compoundwas prepared as an off-white solid (0.045 g, 55%). MS (ES+) m/e 457.2[M+H]⁺.

EXAMPLE 7 Preparation of(S)-N-(1-Formyl-2-phenylethyl)-2-(2-pyridinylethynyl)-4-guinolinecarboxamide

(a)(S)-N-(1-Hydroxymethyl-2-phenylethyl)-2-(2-pyridinylethynyl]-4-quinoline-carboxamide

To a solution of the compound of Example 5(a) (1 g, 2.94 mmol) in dryDMSO (13 mL) under argon atmosphere was added 2-ethynyl pyridine (0.45g, 4.41 mmol; ICN Chemical Company), diphenylphosphine palladiumdichloride (41 mg, 0.059 mmol), copper iodide (22 mg, 0.11 mmol),followed by triethylamine (0.82 mL, 5.88 mmol). The resulting mixturewas heated at 60° C. for 4 h. Methylene chloride (50 mL) was added andthe organic layer was washed with H₂ O, brine, dried (MgSO₄), filtered,and concentrated in vacuo to give an amber oil. The resulting crude oilwas triturated with methylene chloride (40 mL) and methanol (40 mL) andfiltered to yield the title compound as an off-white solid (0.4 g, 33%).MS (ES+) m/e 408.2 [M+H]⁺.

b)(S)-N-(1-Formyl-2-phenylethyl)-2-(2-pyridinylethynyl)-4-quinolinecarboxamide

Following the procedure of Example 1(b) except substituting the compoundof Example 1(a) with the compound of Example 7(a), the title compoundwas prepared as a white solid (0.13 g, 40%). MS (ES+) m/e 406.2 [M+H]⁺.

EXAMPLE 8 Preparation of(S)-N-(1-Formyl-2-phenylethyl)-2-(4-phenyl-1-piperazinyl)-4-quinolinecarboxamide

(a)(S)-N-(1-Hydroxymethyl-2-phenylethyl)-2-(4-phenyl-1-piperazinyl)-4-quinolinecarboxamide

The compound of Example 5(a) (0.1 g, 0.29 mmol) was dissolved in1-phenyl-piperazine (1 mL, 6.5 mmol). The resulting mixture was heatedat 100° C. for 24 h. Methylene chloride (50 mL) was added and theorganic layer was washed with 1N citric acid, saturated NaHCO₃, H₂ O,brine, dried (MgSO₄), filtered, and concentrated in vacuo to give anamber oil. The oil was purified by flash chromatography (silica gel,25-70% EtOAc/hexane) to yield the title compound as a golden yellowsolid (0.03 g, 30%). MS (ES+) m/e 467.4 [M+H]⁺.

b)(S)-N-(1-Formyl-2-phenylethyl)-2-(4-phenyl-1-piperazinyl)-4-quinolinecarboxamide

Following the procedure of Exampie 1(b) except substituting the compoundof Example 1(a) with the compound of Example 8(a), the title compoundwas prepared as a light yellow solid (0.13 g, 40%). MS (ES+) m/e 465.3[M+H]⁺.

EXAMPLE 9 Preparation of(S)-N-(1-Formyl2-phenylethyl)-2-[4-(3-pyridinyl)phenyl]-4-quinolinecarboxamide

(a)(S)-N-(1-Hydroxymethyl-2-phenylethyl)-2-(4-bromophenyl)4quinolinecarboxamide

To a solution of tris(dibenzylideneacetone)dipalladium (0)-chloroformadduct (18 mg, 3 mol %) in toluene (4 mL) under argon atmosphere wasadded triphenylphosphine (19 mg; 1 mol %). The resulting mixture wasstirred at room temperature for 10 min. To the resulting mixture wereadded the compound of Example 5(a) (0.2 g, 0.58 mmol),4-bromophenylboronic acid (0.14 g, 0.7 mmol; Lancaster ChemicalCompany), sodium carbonate (2M solution in H₂ O, 0.6 mL) and ethanol(0.2 mL). The resulting mixture was heated at 90° C. for 14 h. Methylenechloride (50 mL) was added and the organic layer was washed with H₂ O,brine, dried (MgSO₄), filered, and concentrted in vacuo to give an oil.The oil was purified by flash chromatography (silica gel, 25-70%EtOAc/hexane) to yield the title compound as an oil (0.165 g, 61%). MS(ES+) m/e 461.1 [M+H]⁺.

(b)(S)-N-(1-Hydroxymethyl-2-phenylethyl)-2-[4-(3-pyridinyl)phenyl]-4-quinoline-carboxamide

To a solution of the compound of Example 9(a) (0.16 g; 0.35 mmol) intoluene (10 mL) was added 3-pyridyltributyltin (0.158 g; 0.43 mmol;Maybridge Chemical Company) followed by tetrakis(triphenylphosphine)palladium(0) (44 mg, 12 mol %). The resulting mixture was heated at 60°C. for 15 h. Methylene chloride (50 mL) was added and the organic layerwas washed with H₂ O, brine, dried (MgSO₄), filtered, and concentratedin vacuo to give an oil. The oil was purified by flash chromatography(silica gel, 0-15% methanol/CH₂ Cl₂) to yield the title compound as afoam (0.055 g, 30%). MS (ES+) m/e 460.3 [M+H]⁺.

(c)(S)-N-(1-Formyl-2-phenylethyl)-2-[4-(3-pyridinyl)phenyl]-4-quinolinecarboxamide

Following the procedure of Example 1(b) except substituting the compoundof Example 1(a) with the compound of Example 9(b), the title compoundwas prepared as a light yellow solid (0.024 g, 40%). MS (ES+) m/e 458.3{M+H]⁺, 490.3 [M+H+CH₃ OH]⁺.

EXAMPLE 10 Preparation of(S)-N-[1-Formyl-5-[(phenylsulfonyl)amino]pentyl]-2-phenyl-4-guinolinecarboxamide

(a)(S)-N-[1-Carbomethoxy-5-(tetrabutoxyamino]pentyl)]-2-phenyl-4-quinoline-carboxamide

To a solution of 2-phenyl-4-quinoline-carboxylic acid (2 g, 8 mmol,Aldrich Chemical Co.) in methylene chloride (10 mL) was addedbenzotriazol-1-yloxytris-(dimethylamino)phosphoniumhexafluorophosphate(BOP) (3.7 g, 8.4 mmol). The resulting mixture was stirred at roomtemperature for 5 min. (L)-H-(Boc)-Lysine-methyl ester hydrochloride(2.37 g, 8 mmol) was added along with triethylamine (1.2 mL, 8.6 mmol).The resulting mixture was shaken at room temperature for 24 h. Methylenechloride (50 mL) was added and the organic layer was washed with NaHCO₃,H₂ O, brine, dried (MgSO₄), filtered, and concentrated in vacuo to givean oil. The oil was purified by flash chromatography (silica gel, 30-50%EtOAc/hexane) to yield the title compound as a cream solid (3.15 g,80%). MS (ES+) m/e 492.3 [M+H]⁺.

(b) (S)-N-(1-Carbomethoxy)-5-aminopentyl-2-phenyl-4-quinolinecarboxamide

To a cooled solution of the compound of Example 10(a) (0.45 g, 0.92mmol) in dioxane (1 mL) was added a solution of 4N hydrochloric acid indioxane (2.5 mL). The resulting mixture was stirred at room temperaturefor 10 min. The mixture was concentrated in vacuo to give an amber oil.The resulting crude oil was triturated with methylene chloride (40 mL)and methanol (40 mL), diethyl ether (40 mL), and filtered to yield thetitle compound as an off-white solid (0.3 g, 84%). MS (ES+) m/e 392.3[M+H]⁺.

(c)(S)-N-(1-Carbomethoxy)-5-[(phenylsulfonyl)amino]pentyl]-2-phenyl-4-quinoline-carboxamide

To a cooled solution of the compound of Example 10(b) (0.28 g, 0.6 mmol)in THF (9 mL) under argon atmosphere was added N-methylmorpholine (0.23mL, 2.1 mmol) followed by phenyl sulfonyl chloride (0.11 mL, 0.9 mmol).The resulting mixture was slowly warmed to room temperature and wasstirred at room temperature for 16 h. Ice (10 mL) and H₂ O (10 mL) wereadded and the mixture was acidified to pH˜3 with 10% aqueoushydrochloric acid. Methylene chloride (50 mL) was added and the organiclayer was washed with H₂ O, brine, dried (MgSO₄), filtered, andconcentrated in vacuo to give an oil. The oil was purified by flashchromatography (silica gel, 25-60% EtOAc/hexane) to yield the titlecompound as a glassy foam (0.3 g, 94%). MS (ES+) m/e 532.2 [M+H]⁺.

(d)(S)-N-(1-Hydroxymethyl)-5-[(phenylsulfonyl)amino]pentyl]-2-phenyl-4quinoline-carboxamide

To a cooled solution of the compound of Example 10(c) (0.15 g, 0.28mmol) in THF (5 mL) under argon atmosphere was added lithium borohydride(0.21 mL, 0.42 mmol; 2M solution in THF). The resulting mixture wasstirred at room temperature for 16 h. Methylene chloride (2 mL) wasadded and the organic layer was washed with 1N citric acid, H₂ O, brine,dried (MgSO₄), filtered, and concentrated in vacuo to give an amber oil.The oil was purified by flash chromatography (silica gel, 50-80%EtOAc/hexane) to yield the title compound as a glassy foam (0.11 g,83%). MS (ES+) m/e 504.3 [M+H]⁺.

(e)(S)-N-[1-Formyl-5-[(phenylsulfonyl)amino]pentyl]-2-phenyl-4-quinolinecarboxamide

Following the procedure of Example 1 (b) except substituting for thecompound of Example 1(a) with the compound of Example 10(d), the titlecompound was prepared as an off-white solid (0.033 g, 35%). MS (ES+) m/e502.3 [M+H]⁺.

EXAMPLE 11 Preparation of(S)-N-(1-Formyl-5-[(4-pyridylmethoxy)carbonylamino]pentyl]-2-phenyl-4-quinolinecarboxamidedihydrochloride salt

(a)(S)-N-[1-Carbometboxy-5-isocyanopentyl-2-phenyl-4-quinolinecarboxamide

To a cooled solution of the compound of Example 10(b) (1 g, 2.34 mmol)in methylene chloride (10 mL) was added pyridine (0.76 ml, 9.36 mmol)followed by slow addition of phosgene (1.56 mL, 3 mmol; 1.93 M solutionin toluene). The resulting mixture was stirred at 0° C. for 2 h. Theresulting mixture was poured into 0.5 N HCl (25 mL) and ice (15 mL). Theorganic layer was washed with 0.5 N HCl (25 mL) and ice (15 mL). Theaqueous layers were extracted with methylene chloride (40 mL) and thecombined organic layers were washed with brine, dried (MgSO₄), filteredand concentrated to give an oil (0.81 g, 80%) that was used in the nextstep without purification. MS (ES+) m/e 418.3 [M+H]⁺.

(b)(S)-N-(1-Carbomethoxy-5-[(4-pyridylmethoxy)carbonylamino]pentyl]-2-phenyl-4-quinolinecarboxamide

To a solution of the compound of Example 11 (a) (0.81 g, 1.95 mmol)dissolved in toluene (5 mL) was added 4-pyidine carbinol (0.21 g, 1.95mmol; Aldrich Chemical Company). The resulting mixture was refluxed for24 h. Methylene chloride (20 mL) was added and the organic layer waswashed with H₂ O, brine, dried (MgSO₄), filtered, and concentrated invacuo to give an oil. The oil was purified by flash chromatography(silica gel, 0-10% methanol/CH₂ Cl₂) to yield the title compound as aglassy foam (0.46 g, 41%). MS (ES+) m/e 527.3 [M+H]⁺.

(c)(S)-N-(1-Hydroxymethyl)-5-[(4-pyridylmethoxy)carbonylamino]pentyl]-2-phenyl-4-quinolinecarboxamide

Following the procedure of Example 10(d) except substituting for thecompound of Example 10(c) with the compound of Example 11(b), the titlecompound was prepared as an off-white solid (0.14 g, 35%). MS (ES+) m/e499.3 [M+H]⁺.

(d)(S)-N-(1-Formyl-5-[(4-pyridylmethoxy)carbonylamino]pentyl]-2-phenyl-4-quinolinecarboxamidedihydrochloride salt

Following the procedure of Example 1(b) except substituting for thecompound of Example 1(a) with the compound of Example 11(c), the titlecompound was prepared as a light yellow solid. The solid was dissolvedin ethanol (1 mL) and ethereal HCl (2 mL) was added. The precipitatedsolid was filtered and dried in vacuo to yield the title compound as anoff-white solid (0.03 g, 35%). MS (ES+) m/e 497.3 [M+H]⁺, 529.3 [M+H+CH₃OH]⁺.

EXAMPLE 12 Preparation of(S)-N-(1-formyl-2-(phenylethyl)-2-(phenylethynyl)-4-quinoline-carboxamide

(a)(S)-N-(1-hydroxymethyl-2-(phenylethyl)-2-(phenylethynyl)4-quinoline-carboxamide

To a solution of the compound of Example 5(a) (4.4 g, 12.9 mmol) in dryDMSO (50 mL) under argon atmosphere was added phenylacetylene (2.13 mL,19.4 mmol; Aldrich Chemical Company), diphenylphosphine palladiumdichloride (181 mg, 2% mol), copper iodide (98 mg, 4% mol), followed bytriethylamine (3.6 mL, 25.8 mmol). The resulting mixture was heated at60° C. for 4 h. Methylene chloride (50 mL) was added and the organiclayer was washed with H₂ O, brine, dried (MgSO₄), filtered, andconcentrated in vacuo to give an amber oil. The resulting crude oil wastriturated with methylene chloride (40 mL) and methanol (40 mL) andfiltered to yield the title compound as an off-white solid (4.4 g, 60%).MS (ES+) m/e 407.2 [M+H]⁺.

(b)(S)-N-(1-formyl-2-(phenylethyl)-2-(phenylethynyl)-4-quinolinecarboxamide

Following the procedure of Example 1(b) except substituting the compoundof Example 1(a) with the compound of Example 12(a), the title compoundwas prepared as a white solid (1.67 g, 50%). MS (ES+) m/e 405.2 [M+H]⁺.

EXAMPLE 13 Preparation ofN-[3-(n-butylamino)-2,3-dioxo-1-(phenylmethyl)]-2-(phenylethynyl)-4-quinolilnecarboxamide

(a) (S)-N-(1-formyl-2-phenylethyl)-2-chloro-4-quinolinecarboxamide

Following the procedure of Example 1(b) except substituting(S)-N-(1-hydroxymethyl-2-phenylethyl)-2-phenyl-4-quinolinecarboxamidewith(S)-N-(1-hydroxymethyl-2-phenylethyl)-2-chloro-4-quinolinecarboxamide,the title compound was prepared as a white solid (1.1 g, 50%). MS (ES+)m/e 339 [M+H]⁺.

(b)(S)-N-3-[(n-butylamino)-2-hydroxy-3-oxo-1-(phenylmethyl)propyl]-2-(chloro)-4-quinolinecarboxamide

To a cooled solution (0° C.) of TiCl₄ (1.55 mL, 1.55 mmol; 1M solutionin CH₂ Cl₂) in methylene chloride (5 mL) was added n-butyl isocyanide(162 μL, 1.5 mmol). The resulting mixture was stirred at 0° C. for 3 h.The resulting solution was cooled to -78° C. and(S)-N-(1-formyl-2-phenylethyl)-2-chloro-4-quinolinecarboxamide (0.5 g,1.47 mmol) was added. The resulting mixture was warmed to roomtemperature over 1 h and stirred at room temperature for 24 h. To themixture was added 1N HCl (2 mL) and stirred for 30 min. The mixture wasdiluted with EtOAc and the aqueous layer basified with NaOH andextracted with EtOAc. The combined organic layer was washed with brineand dried over Na₂ SO₄. TLC (40-60 EtOAc:Hexane) showed thedisappearance of starting material and appearance of faster and slowerrunning spots. Solvent was removed in vacuo and the resulting mixturewas triturated with ether to give the desired compound as precipitatedwhite solid (0.2g, 35%). MS (ES+) m/e 440 [M+H]⁺.

(c)(S-N-[3-(n-butylamino)-2-hydroxy-3-oxo-1-(phenylmethyl)propyl]-2-(phenyl-ethynyl)-4-quinolinecarboxatuide

Following the procedure of Example 12(a) except substituting thecompound of Example 5(a) with(S)-N-[3-(n-butylamino)-2-hydroxy-3-oxo-1-(phenylmethyl)propyl]-2-(chloro)4-quinolinecarboxamide,the title compound was prepared as an off-white solid (65 mg, 50%). MS(ES+) m/e 506 [M+H]⁺.

(d)N-[3-(n-butylamino)-2,3-dioxo-1-(phenylmethyl)]-2-(phenylethynyl)4-quinolinecarboxamide

Following the procedure of Example 1(b) except substituting the compound(S)-N-(1-hydroxymethyl-2-phenylethyl)-2-phenyl-4-quinollinecarboxamidewith(S)-N-[3-(n-butylamino)-2-hydroxy-3-oxo-1-(phenylmethyl)propyl]-2-(phenylethynyl)-4-quinolinecarboxamide,the title compound was prepared as a white solid (45 mg, 50%). MS (ES+)m/e 504.2 [M+H]⁺.

EXAMPLE 14

    ______________________________________                                        Ingredients             Mg./Capsule                                           ______________________________________                                        (S)--N-(1-formyl-2-phenylethyl)-2-pyridinyl-                                                          250.00                                                ethynyl-4-quinoline-carboxamide                                               Magnesium Stearate       5.00                                                 Lactose                 100.00                                                ______________________________________                                    

The ingredients are thoroughly mixed and filled into a hard gelatincapsule.

EXAMPLE 15

    ______________________________________                                        Ingredients              Mg./Tablet                                           ______________________________________                                        (S)-2-[1,1'-biphenyl]-2-yl-N-(1-formyl-2-phenyl-                                                       100.00                                               ethyl)-4-quinolinecarboxamide                                                 Lactose                  250.00                                               Starch                    13.00                                               Talc                      5.00                                                Magnesium Stearate        2.50                                                ______________________________________                                    

The lactose and quinolinecarboxamide are mixed and granulated with hot10% gelatin. The granules are dried and passed through a #20 meshscreen. The granules are then mixed with the starch, talc and magnesiumstearate and compressed into a tablet.

One tablet is administered four times a day to mammals for treatment ofneurodegenerative diseases.

EXAMPLE 16

    ______________________________________                                        Ingredients             Amounts/Mg.                                           ______________________________________                                        (S)--N-(1-formyl-2-phenylethyl)-2-pyridinyl-                                                           75.00                                                ethynyl-4-quinolinecarboxamide                                                DMSO                    500.00                                                Sodium Chloride         375.00                                                Sodium Bisulfite        100.00                                                Water for Injection q.s.                                                                              100 ml                                                ______________________________________                                    

The quinolinecarboxamide is dissolved in the DMSO and 50% of the water.The salts are thoroughly dissolved and the volume is brouht up to 100ml. The solution is then filtered and filled into ampules andautoclaved.

EXAMPLE 17

    ______________________________________                                        Ingredients               Amounts/Mg.                                         ______________________________________                                        (S)-2-[1,1'-biphenyl]-2-yl-N-(1-formyl-2-phenyl-                                                        150.00                                              ethyl)-4-quinolinecarboxamide                                                 Peanut Oil                300.00                                              ______________________________________                                    

The ingredients are mixed to a thick slurry and filled into soft gelatincapsules. One capsule is administered orally to mammals for treatment ofneurodegenerative diseases.

What is claimed is:
 1. A compound of the formula: ##STR14## in which: Xis CH or N;R is CH₂ Ph, --(CH₂)₃ CH₂ NR₁ R₃, CH₂ CH(CH₃)₂ or CH₂ PhOR₂ ;R₁ is COOCH₂ Ph, SO₂ CH₃, SO₂ aryl, COOCH_(2pyridyl) (or substitutedpyridyl); R₂ is H, CH₃,CH₂ Ph or CH_(2pyridyl) ; R₃ is H, CH₃ or loweralkyl; Z is CHO, COCH₂ F, COCOOH, COCOOalkyl, COCONHalkyl, COCO(CH₂)_(n)aryl, COCONHCH(R)COOH or COCH₂ O-3-(phenylisoxazol-5-yl); n is 1 to 6;##STR15## Y=absent, phenyl, substituted phenyl, pyridyl or substitutedpyridyl, or a pharmaceutically acceptable salt thereof.
 2. A compound ofclaim 1 in which the C-5 sterochemistry is S.
 3. A compound of claim 2in which X is N.
 4. A compound of claim 3 in which Z is CHO.
 5. Acompound of claim 4 in which R is CH₂ Ph or --(CH₂)₃ CH₂ NHR₁.
 6. Acompound of claim 5 being(S)-N-(1-formyl-2-phenylethyl)-2-phenyl-4-quinolinecarboxamide.
 7. Acompound of claim 5 being(S)-2-(4-chlorophenyl-N-(1-formyl-2-phenylethyl)-4-quinolinecarboxamide.8. A compound of claim 5 being(S)-2-[1,1'-biphenyl]-4-yl-N-(1-formyl-2-phenylethyl)-4-quinolinecarboxamide.9. A compound of claim 5 being(S)-2-(1-adamantyl)-N-(1-formyl-2-phenylethyl)-4-quinolinecarboxamide.10. A compound of claim 5 being(S)-N-(1-formyl-2-phenylethyl)-2-(4-phenoxyphenyl-4-quinolinecarboxamide.11. A compound of claim 5 being(S)-2-[1,1'-biphenyl]-2-yl-N-(1-formyl-2-phenylethyl)-4-quinolinecarboxamide.12. A compound of claim 5 being(S)-N-(1-formyl-2-phenylethyl)-2-(2-pyridinylethynyl)-4-quinolinecarboxamide.13. A compound of claim 5 being(S)-N-(1-formyl-2-phenylethyl-2-(4-phenyl-1-piperazinyl)-4-quinolinecarboxamide.14. A compound of claim 5 being(S)-N-[1-formyl-2-phenylethyl]-2-[(3-pyridinyl)-4-phenyl]-4-quinolinecarboxamide.15. A compound of claim 5 being(S)-N-[1-formyl-5-[(phenylsulfonyl)amino]-pentyl]-2-phenyl4-quinolinecarboxamide.16. A compound of claim 5 being(S)-N-[1-formyl-5-[N'-(carbo-4-pyridinemethyloxy)pentyl]-2-[phenyl]-4-quinolinecarboxamide.17. A compound of claim 5 being(S)-N-(1-formyl-2-(phenylethyl)-2-(phenylethynyl)-4-quinolinecarboxamide.18. A compound of claim 5 beingN-[3-(n-butylamino)-2,3-dioxo-1-(phenylmethyl)]-2-phenylethynyl)-4-quinolinecarboxamide.19. A pharmaceutical composition in dosage unit form for inhibitingcalpain comprising a pharmaceutical carrier and an effective amount ofthe compound as described in claim
 1. 20. A method of inhibiting calpainwhich comprises administering to an animal or human in an amountsufficient to inhibit calpain a compound as described in claim
 1. 21. Amethod of treating neurodegenerative diseases which comprisesadministering to an animal or human in need thereof orally or byinjection a sufficient amount of a compound of claim
 1. 22. The methodof claim 20 wherein the amount is from about 50 to about 500 mg of thecompound per dosage unit and the administration is orally.
 23. Themethod of claim 20 wherein the amount is from about 0.1 to 140 mg/kg ofbody weight of the animal or human and the administration isparenterally.